Multiple monohydroxylation products from rac-camphor by marine fungus Botryosphaeria sp. Isolated from marine alga Bostrychia radicans

Article abstract of DOI:10.21577/0103-5053.20160262

This manuscript describes the biooxidation of rac-camphor using whole cells of marine-derived fungus Botryosphaeria sp. CBMAI 1197. The main biotransformation products of this monoterpene were achieved via a hydroxylation reaction and occurred with 5 days of rac-camphor incubation. Products were identified by means of gas chromatography mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) data. The major hydroxylated products were 6-endo-hydroxycamphor, 6-exo-hydroxycamphor, 5-exo-hydroxycamphor, 5-endo-hydroxycamphor, 3-exo-hydroxycamphor and 8-hydroxycamphor. The 6-exo-hydroxycamphor was obtained through a retro-aldol reaction when 6-endo-hydroxycamphor was maintained in presence of CDCl₃; this isomerization was confirmed by ¹H NMR and GC-MS data.

Full text of DOI:10.21577/0103-5053.20160262

http://dx.doi.org/10.21577/0103-5053.20160262
J. Braz. Chem. Soc., Vol. 28, No. 3, 498-504, 2017.
Printed in Brazil - ©2017 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00
Short Report
Multiple Monohydroxylation Products from rac-Camphor by Marine Fungus
Botryosphaeria sp. Isolated from Marine Alga Bostrychia radicans
Hugo C. R. de Jesus,a,b,c Alex H. Jeller,d Hosana M. Debonsi,e Péricles B. Alvesb and
André L. M. Porto*,a
aLaboratório de Química Orgânica e Biocatálise, Instituto de Química de São Carlos,
Universidade de São Paulo, Av. João Dagnone, 1100, Ed. Química Ambiental, J. Santa Angelina,
13563-120 São Carlos-SP, Brazil
bDepartamento de Química, Universidade Federal de Sergipe, São Cristóvão,
49100-000 Aracajú-SE, Brazil
cLaboratório Dalton de Espectrometria de Massas, Instituto de Química, Universidade Estadual de
Campinas (UNICAMP), 13803-862 Campinas-SP, Brazil
dCentro de Estudos em Recursos Naturais, Universidade Estadual de Mato Grosso do Sul,
Rod. Itahum Km 12, s/n, 79804-970 Dourados-MS, Brazil
eDepartamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto,
Universidade de São Paulo, Via do Café s/n, 14040-903 Ribeirão Preto-SP, Brazil
This manuscript describes the biooxidation of rac-camphor using whole cells of marine-derived
fungus Botryosphaeria sp. CBMAI 1197. The main biotransformation products of this monoterpene
were achieved via a hydroxylation reaction and occurred with 5 days of rac-camphor incubation.
Products were identified by means of gas chromatography mass spectrometry (GC-MS) and nuclear
magnetic resonance (NMR) data. The major hydroxylated products were 6-endo-hydroxycamphor,
6-exo-hydroxycamphor, 5-exo-hydroxycamphor, 5-endo-hydroxycamphor, 3-exo-hydroxycamphor
and 8-hydroxycamphor. The 6-exo-hydroxycamphor was obtained through a retro-aldol reaction
when 6-endo-hydroxycamphor was maintained in presence of CDCl3; this isomerization was
confirmed by 1H NMR and GC-MS data.
Keywords: terpene, hydroxylation, biocatalysis, marine endophytic fungi
Introduction
pollinators.1 This class of compounds has been widely
used for bactericidal, virucidal, fungicidal, antiparasitical,
insecticidal, medicinal and cosmetic applications especially
in pharmaceutical, sanitary, cosmetic, agricultural and food
industries.5,6
Considering the wide variety of applications and
activities of monoterpenes, there are attempts to modify
its basic structure to improve its properties. This can be
accomplished by chemical synthesis and biotransformation
reactions. However, organic synthesis is not typically
satisfactory for performing structural modifications of
terpenoids because of its low reactivity. This produces
substances in poor yields.
The mono and sesquiterpenes belong to the largest
class of secondary metabolites. They are known not
only as raw materials for flavor and fragrance, but also
as biologically active substances.1 The (−)-camphor
is a bicyclic monoterpene and is one of the oldest
known organic compounds. It is a major constituent of
essence of dalmatian sage (Salvia officinalis).2,3 It has a
camphoraceous odor and is used commercially as a moth
repellent and in skin care formulations for cosmetic and
pharmaceutical purposes.4
Monoterpenes in plants have mainly ecological roles.
They repel herbivores, minimize fungus, and attract
The oxidation of inactivated C–H bonds under
mild conditions has no equivalent in classical synthetic
methodologies; P450 enzymes also have significant
*e-mail: almporto@iqsc.usp.br

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499
potential in biotransformation applications. For example,
cytochrome P450 from Pseudomonas putida catalyzed the
selective oxidation of camphor to 5-exo-hydroxycamphor.7
There are few articles in the literature describing
monohydroxylation of camphor. Nakahashi and Miyazawa8
used Salmonella typhimurium OY1000/2A6 of human
cytochrome P450, employing the cofactor NADPH-P450
reductase. This hydroxylated the (−)-5-exo-hydroxycamphor
and (−)-8-hydroxycamphor. Miyazawa and Miyamoto9 used
the larvae of the common cutworm (Spodoptera litura) to
biotransform rac-camphor to 5-endo-hydroxycamphor,
5-exo-hydroxycamphor (5) and 8-hydroxycamphor.
However, to the best of our knowledge, there is not report
on camphor biotransformation by marine fungi.
The mass spectrometry (MS) used electron capture
detection, electronic impact of 70 eV, scanning speed
of 1,000 Da sec-1, scan interval of 0.50 fragments s-1
and fragments detected from 40-500 Da. The 1H and 13C
nuclear magnetic resonance (NMR) spectra were obtained
on a Bruker/AC-200 spectrometer (1H at 200 MHz and
13C at 50 MHz). The spectra were obtained in deuterium
chloroform (CDCl3), and the chemical shifts are given
in ppm with tetramethylsilane (TMS) as the internal
standard.
Isolation, identification and preparation of stock cultures
of marine-derived fungus Botryosphaeria sp. CBMAI 1197
We recently investigated the bio-oxidation of
natural products using marine-derived fungi.10 In this
study, we used whole cells of Brazilian marine fungus
Botryosphaeria sp. CBMAI 1197 in the biohydroxylation
reaction of monoterpene rac-camphor (1). The fungus
Botryosphaeria sp. CBMAI 1197 was isolated from red
marine alga Bostrychia radicans.11 Therefore, different
hydroxylated compounds were produced by fungal
bioconversion of monoterpene.
The Botryosphaeria sp. CBMAI 1197 fungus strain
was isolated from red marine alga Bostrychia radicans
collected in Ubatuba in the South Atlantic Ocean off
the northern coast of the state of São Paulo, Brazil, in
September 2007 by Prof Hosana M. Debonsi (FCFRP/
USP).11 This alga was identified by conventional taxonomic
methods by N. S. Yokoya (Botanical Institute of São Paulo,
IBOT/SP, Brazil). A voucher specimen was deposited at
the Herbarium of Botanical Institute of São Paulo, Brazil,
under accession number SP 365678.
Experimental
Stock cultures of the marine-derived fungus was
stored in Petri dishes containing a 2% malt solid culture
medium based on artificial sea water (ASW) [CaCl2·2H2O
(1.36 g L−1), MgCl2·6H2O (9.68 g L−1), KCl (0.61 g L−1),
NaCl (30.0 g L−1), Na2HPO4 (0.014 mg L−1), Na2SO4
(3.47 g L−1), NaHCO3 (0.17 g L−1), KBr (0.1 g L−1),
SrCl2·6H2O (0.040 g L−1), H3BO3 (0.030 g L−1)], agar
(20 g L−1), and malt extract (20 g L−1). The pH of the
medium was adjusted to 8 with 3 mol L-1 KOH. The Petri
dishes were maintained at 4 °C for later use.
General methods
The rac-camphor (1) (> 96%) used in the
biotransformation experiments was purchased from Sigma-
Aldrich. Technal T-421 and Superohm orbital shakers were
employed in the biocatalytic transformation experiments.
Sterile materials were used for the biotransformation
experiments, and Botryosphaeria sp. CBMAI 1197 was
handled in a Veco biological safety cabinet. Thin-layer
chromatography (TLC) was performed on pre-coated
plates (aluminum foil, silica gel 60 F254 Sorbent, 0.25 mm,
from Merck).
Biotransformation of rac-camphor (1) by marine-derived
fungus Botryosphaeria sp. CBMAI 1197
Enzymatic products were purified by column
chromatography (CC) on silica gel (SiO2; 230-400 mesh,
from Across) eluted with n-hexane/EtOAc mixtures in
increasing polarity degrees. The products ofrac-camphor (1)
analyses were carried out using Shimadzu 2010
GC/FID equipped with an auto-injector AOC20i and
DB-5MS (30 m × 0.25, 0.25 µm) column. The injector
and detector temperature were maintained at 250 °C, the
split ratio of the injector was 1:20, the carrier gas was He
at 88.2 kPa, and the flux was 1.5 mL min-1. The program
used for gas chromatography (GC) analysis was as follows:
initial temperature of 50 °C, final temperature of 232 °C,
ramp rate of 7 °C, and final time of 30 min.
The fungus was cultured in 250 mL Erlenmeyer flasks
containing 100 mL of liquid culture medium with 2% malt
extract (3 days, at 32 °C) on an orbital shaker (150 rpm)
in ASW. After this time, the rac-camphor (1) (50.0 mg)
previously dissolved in 400 µL of dimethyl sulfoxide was
added and mixed into the liquid culture medium. The
biocatalytic reactions were analyzed periodically (24,
48, 72, and 120 h) by collecting 2 mL of samples and
adding 2 mL of ethyl acetate with vortex mixing and then
centrifuged at 6000 rpm for 6.0 min in a HERMLE Z-200 A.
The organic phase was analyzed by GC-FID and GC-MS.
The experiments were conducted in triplicate. After 120 h,
the biotransformation products were isolated.

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Multiple Monohydroxylation Products from rac-Camphor by Marine Fungus Botryosphaeria sp.
J. Braz. Chem. Soc.
Purification of hydroxylated products obtained by
1197 fungus. This demonstrated that products 2-7 formed at
differentreactiontimes(24,72and120h)andweremaintained
by the same chromatographic profile (had no change) during
the biotransformation process (Figure 1). A control stability
experiment was performed in which the liquid culture
medium (absent of fungus) was added to therac-camphor (1);
the compound was stable for at least 120 h (Figure 1).
Several compounds may undergo spontaneous reactions.
These are sometimes mistakenly associated with enzymatic
conversions. Thus, rac-camphor (1) was added to the culture
medium in the absence of fungus to study stability. The
compound remained intact. Thus, the peaks in Figure 1 are
products 2-7 due to biotransformation of rac-camphor (1) by
enzymes from Botryosphaeria sp. CBMAI 1197.
In the biotransformation of rac-camphor (1) by
Botryosphaeria sp. CBMAI 1197 were obtained six main
metabolites 2-7, which were previously identified by mass
spectrometry data. In addition, four substances (3, 5-7)
were isolated in mixture by chromatographic column and
identified by NMR and MS analyses. All of the data were
compared with those described in the literature (Scheme 1).
MS analysis of metabolites 2-7 showed a specific
molecular ion peak (M+•) at m/z 168, indicating the presence
of a hydroxyl group added to the substrate (152 g mol-1) by
Botryosphaeria sp. CBMAI 1197 (Figure 2).
Figure 3 shows the chromatogram (GC-MS)
fractions (A-D) containing compounds 3-7 produced by
biotransformation. The compounds were partially purified
through normal phase chromatography column. These
fractions showed the oxidized products 3-6. Compounds 3
and 5 were isolated with 81 and 75% purity, respectively,
and were identified by NMR analyses (Figures 3A-B).
Metabolites 6 and 7 were obtained as a mixture. These
molecules were structurally identified by NMR data
(Figures 3C-D). The fraction obtained by chromatographic
separation and analyzed by GC-MS showed compound 3
in a mixture of substances 5-6 (Figure 3C). Therefore,
biotransformation of rac-camphor (1)
The isolation of the hydroxylated products was
performed using CC; with stationary phase flash silica gel
60 (230-400 mesh) from Across. The separation progress
was monitored by TLC on pre-coated silica gel 60 F254
from Sorbent Technologies. The samples were observed
under UV light (254 nm) and developed with anisaldehyde
solution (100 mL of acetic acid, 1 mL of concentrated
sulfuric acid and 1 mL of p-anisaldehyde) or alcoholic
solution of vanillin (125 mL ethanol, 25 mL of concentrated
sulfuric acid and 6 g vanillin). Hexane and ethyl acetate
was used as the mobile phase for elution of components.
This started with 100% hexane (175 mL) and the polarity
gradually increased: 9.5:0.5 (75 mL), 9.0:1.0 (75 mL),
8.5:1.5 (75 mL), 8.0:2.0 (150 mL), 7.5:2.5 (200 mL),
7.0:3.0 (200 mL), and 6.5:3.5 (100 mL).
Results and Discussion
Biotransformation of rac-camphor (1) by the whole cells
of marine-derived fungus Botryosphaeria sp. CBMAI 1197
Botryosphaeria sp. CBMAI 1197 transformed the
rac-camphor (1) to different hydroxylated products 2-7.
The hydroxylation mediated by Botryosphaeria genus has
been described in the literature in different natural products
including biooxidation reactions of 3-keto-bisnorcholeno,
nootkatone, ambrox, sclareol, sclareolide and valencene.12-14
We initially carried out a kinetic study of the
biotransformation reaction of rac-camphor (1) up to 120 h.
After this period, the chromatographic profile obtained by
GC-flame ionization detection (FID) analysis was unchanged
indicating that the biotransformation was stabilized in these
conditions. The biotransformation of rac-camphor (1) was
mediated by whole cells from Botryosphaeria sp. CBMAI
Figure 1. GC-FID analyses of biotransformation reaction of rac-camphor (1) by marine endophytic fungus Botryosphaeria sp. CBMAI 1197 in different
times. Control: liquid culture medium plus rac-camphor (1) in fungus absence.

Vol. 28, No. 3, 2017
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501
metabolite 6 was in mixture of compound 3 and 5.
Compound 3 (majority in fraction C) was obtained in
mixtures with metabolites 5-7. Compound 7 was identified
in this mixture and could be seen in fraction D (Figure 3D).
In addition, compound 2 was not isolated, and compound 4
was obtained from isomerization of compound 3.
The identification and characterization of hydroxylated
compounds 2-7 are described here. The exact position of the
hydroxyl group was confirmed by NMR and MS analyses
of the substances (even in mixture). These were purified by
flash chromatography column and compared with literature
data.9 These substances obtained via biotransformation of
rac-camphor (1) by Botryosphaeria sp. CBMAI 1197 were
obtained as white solids.
1
The H NMR spectra of the carbinolic hydrogen
region showed that the fractions contained mixture of
metabolites 3-7. This explained the position of the hydroxyl
groups in rac-camphor (1) (Supplementary Information).
Fraction A (Figure 3A) had a peak at d 9.86.
Surprisingly, this signal was not assigned to any hydrogen
Scheme 1. Biohydroxylation of rac-camphor (1) by whole cells of
Botryosphaeria sp. CBMAI 1197 yielding the compounds 2-7.
Figure 2. Mass spectra for the hydroxylated derivative metabolites 2-7 from rac-camphor (1) obtained by Botryosphaeria sp. CBMAI 1197.

502
Multiple Monohydroxylation Products from rac-Camphor by Marine Fungus Botryosphaeria sp.
J. Braz. Chem. Soc.
Figures 3. (A-D) GC-MS (70 eV) chromatograms of the fractions 3A-D containing the compounds 3-7 yield by biotransformation of rac-camphor (1)
using Botryosphaeria sp. CBMAI 1197.
of the hydroxyl-camphor metabolite. The signal at d 9.86
was obtained from an aldehyde intermediate when the
sample of 6-endo-hydroxycamphor (3) was solubilized in
deuterated chloroform. According to Grogan et al.,15 the
6-endo-hydroxycamphor (3) undergoes an isomerization
1
through a retro-aldol reaction confirmed with H NMR
shield at 9.86 ppm. Scheme 2 shows the spontaneous
isomerization mechanism of 6-endo-hydroxycamphor (3)
to obtain the 6-exo-hydroxycamphor (4) (NMR spectrum,
Supplementary Information).
Figure 4. Chromatograms obtained by GC-MS analysis. (A) Spontaneous
isomerization of 6-endo-hydroxycamphor (3) to 6-exo-hydroxycamphor
(4) in presence of CDCl3; (B) 6-endo-hydroxycamphor (3) in absence
of CDCl3.
Scheme 2. Spontaneous isomerization of 6-endo-hydroxycamphor (3) to
6-exo-hydroxycamphor (4) in presence of CDCl3.
hydrogen of 6-endo-hydroxycamphor (3). Therefore, the
structural characterization of hydroxylated metabolite 3 was
carried out by 1H NMR analysis. This showed the presence
of methyl groups at 0.83 ppm (3H, s) and 0.98 ppm (6H, s).
These signals were attributed to the hydrogens at positions 9,
8 and 10 of 6-endo-hydroxy-camphor (3) (Table 1). Also,
the 6-endo-hydroxy-camphor (3) had coupling of 18 Hz
between H-3endo (1.99 ppm) and H-3exo (d 2.35-2.48) for
vicinal hydrogens in this position. At 2.15 ppm, we noted a
triplet of the methynic hydrogen bound in position 4 of the
6-endo-hydroxy-camphor (3). The hydroxyl group assigned
to the position H-6endo was responsible for the signal at
GC-MS analysis after dilution of the sample containing
6-endo-hydroxycamphor (3) in presence of CDCl3 after
16 hours confirmed the existence of isomerization (Figure 4
and Scheme 2). The 6-exo-hydroxycamphor (4) signal
increased and the peak of 6-endo-hydroxycamphor (3)
decreased (Figures 3A and 4). The increased stability of the
6-exo-diastereomer (4) explains the balance shift.
In the 1H NMR spectrum of fraction A (Figure 3A and
Supplementary Information), we noted a double doublet at
dH 4.18 (J = 8.0 and 2.0 Hz) that was attributed to 6-exo-

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Jesus et al.
503
Table 2. 1H NMR data of 5-exo-hydroxy-camphor (5) (200 MHz, CDCl3)
obtained by hydroxylation of rac-camphor (1) using Botryosphaeria sp.
CBMAI 1197
4.18 ppm. This showed a constant coupling at 8.0 and 2.0 Hz.
The complete structural determination of 6-endo-hydroxy-
camphor (3) was confirmed by comparison of 1H NMR data
described in the literature (Table 1).9 Other characteristic
signals can be seen in Table 1.
Experimental data for Literature data for 5-exo-
5-exo-hydroxy-
camphor (5)
hydroxy-camphor (5)9
Table 1. 1H NMR data of 6-endo-hydroxy-camphor (3) (200 MHz, CDCl3)
obtained by hydroxylation of rac-camphor (1) using Botryosphaeria sp.
CBMAI 1197
Position
3-endo
3-exo
4
dH (J in Hz)a / ppm
1.70 (d, 18.0)
2.34 (d, 18.0, 6.0)
2.16 (d, 6.0)
4.02 (dd, 6.0, 4.0)
1.85 (m)
dH (J in Hz)b / ppm
1.70 (d, 18.5)
2.33 (dd, 18.5, 5.0)
2.16 (dd, 5.0, 1.0)
4.02 (dd, 7.5, 3.5)
1.85 (dd, 14.0, 7.5)
1.79 (ddd, 14.0, 3.5, 1.0)
0.85 (s, 6H)
Experimental data for
6-endo-hydroxy-
camphor (3)
Literature data for 6-endo-
hydroxy-
5-endo
6-endo
6-exo
8
camphor (3)9
1.81 (m)
Position
3-endo
3-exo
4
dH (J in Hz)a / ppm
1.99 (d, 18.0)
dH (J in Hz)b / ppm
1.98 (d, 18.5)
0.85 (s, 3H)
9
1.25 (s, 3H)
1,25 (s, 3H)
10
0.93 (s, 3H)
0.93 (s, 3H)
2.35-2.48 (m)
2.43 (ddd, 18.5, 5.0, 3.0)
2.14 (dd, 5.0, 4.5)
1.33 (dd, 13.5, 2.5)
a1H NMR data (200 MHz, CDCl3); b1H NMR data (400 MHz, CDCl3).
2.15 (t, 4.0)
5-endo
1.34 (dd, 14.0, 2.0)
2.54 (dddd, 13.5, 10.0,
4.5, 3)
5-exo
2.50-2.61 (m)
6-exo
8 and 10
9
4.18 (dd, 8.0, 2.0)
0.98 (s, 6H)
4.16 (dd, 10, 2.5)
0.96 (s, 6H)
0.83 (s, 3H)
0.82 (s, 3H)
a1H NMR data (200 MHz, CDCl3); b1H NMR data (400 MHz, CDCl3).
The fraction B chromatogram (Figure 3B) showed
compound 5 as the majority presence. The carbinolic
hydrogen region of the 1H NMR spectrum showed a double
doublet at 4.02 ppm that was attributed to the 5-endo-
hydrogen yielding 5-exo-hydroxycamphor (5). This doublet
had coupling constants of 6.0 and 4.0 Hz indicating an
endo-endo correlation between the hydrogens in positions
H-5 and H-6, and endo-exo between the 5-endo-hydrogen
and 6-exo-hydrogen, respectively. The other signals
were compared with the literature and corroborated the
characterization of 5-exo-hydroxycamphor (5) (Table 2).9
Figure 5. Specific signals obtained by 1H NMR spectrum for carbinolic
hydrogens of monohydroxylated derivatives (3, 5 and 6) of rac-camphor
(1) obtained from fraction C.
there were two doublets at 3.50 ppm (d, J = 12.0 Hz) and
3.74 ppm (d, J = 12.0 Hz). This is usually designated as
the roof effect and was allocated to two diastereotopic
carbinolic hydrogens of 8-hydroxycamphor (7) (Figure 6).
Miyazawa and Miyamoto9 assigned readings on 3.52 ppm
(d, J = 11.0 Hz) and 3.74 ppm (d, J = 11.0 Hz) to hydrogens
of C-8 of 8-hydroxycamphor (7).
Finally, compound 2 was not isolated, and there are ten
sites of hydroxylation in the rac-camphor (1) (Figure 7).
Compound 2 might have been a possible hydroxylated
stereoisomer according to the data of spectrum mass. This
was suggested at position-2 (Figure 2 and Supplementary
Information).
This study showed the camphor biotransformation
by a marine environment fungus. The identification of
monohydroxylated products from the oxidation of camphor
is important for the identification of metabolites or the use
of hydroxylated derivatives of camphor in pharmaceutical
formulations.
1
The H NMR spectrum of fraction C (Figure 3C
and Supplementary Information) showed the presence
of three groups with characteristic signal for carbinolic
hydrogens. The majority signal at 4.18 ppm concerned
the 6-exo-hydrogen of 6-endo-hydroxycamphor (3).
The signal at 4.02 ppm was the 5-endo-hydrogen of
5-exo-hydroxycamphor (5), and the other signal type
multiplet was at 4.58-4.66 ppm and was assigned to 5-exo-
hydrogen of 5-endo-hydroxycamphor (6) (Figure 5). This
characterization agrees with the data obtained by Miyazawa
and Miyamoto9 who proposed substance 6 presents a shield
in 4.64 ppm as a dddd (J = 9.5, 4.5, 4.0 and 2.0 Hz).
Fraction D (Figure 2D and Supplementary Information)
showed hydroxylation at 6-endo for compound 3 (4.18 ppm,
dd, J = 8.0. and 2.0 Hz), 5-exo for compound 5 (d 4.02)
and 5-endo for compound 6 (d 4.58-4.66, m). In addition,

504
Multiple Monohydroxylation Products from rac-Camphor by Marine Fungus Botryosphaeria sp.
J. Braz. Chem. Soc.
Acknowledgments
H. C. R. J. thanks to Coordenação de Aperfeiçoamento
de Pessoal de Nível Superior (CAPES) for scholarships. The
authors thank to Conselho Nacional de Desenvolvimento
Científico e Tecnológico (CNPq) and Fundação de Amparo
à Pesquisa do Estado de São Paulo (FAPESP) for financial
support. A. L. M. P. thanks to 15th Anniversary of the
BIOprospecTA-BIOTA program.
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Conclusions
The Brazilian marine-derived fungus Botryosphaeria sp.
CBMAI 1197 is an efficient biocatalyst that promotes
mono-biohydroxylation of rac-camphor (1). The main
biotransformation product was 6-endo-hydroxycamphor (3)
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We would like to inform that this manuscript presents to
the reader the supplementary information (NMR spectra and
proposal of MS fragmentation for identified compounds),
available free of charge at http://jbcs.sbq.org.br.
Submitted: June 16, 2016
Published online: September 26, 2016
FAPESP has sponsored the publication of this article.